摘要
目的：应用cTnTR141W转基因扩张型心肌病小鼠,研究真武汤对家族遗传性扩张型心肌病心肌细胞结构及cTnTR141W基因表达的影响。方法：（1）利用RT-PCR和HpaⅡ酶切检测空白组正常小鼠和转基因组小鼠cTnTR141W的表达水平。（2）将cTnTR141W转基因小鼠随机分为模型组和真武汤治疗组,1个月后作为给药终点,（1）光镜下观察空白组、模型组、真武汤治疗组心脏结构的病理改变;（2）透射电镜观察各组小鼠心肌的超微结构;（3）HpaⅡ酶切和RT-PCR鉴定各组小鼠心肌组织中cTnTR141W转基因的表达水平。结果：（1）HpaⅡ酶切和RT-PCR鉴定正常小鼠与转基因鼠cTnTR141W的表达结果表明：空白组小鼠的基因经酶切后未检测到转基因cTnTR141W,模型组小鼠的基因经酶切后检测到转基因cTnTR141W,转基因cTnTR141W是内源性cTnT的1倍;（2）扫描切片观察显示,模型组小鼠的心脏大于正常组小鼠的心脏,左室明显扩张,且室壁变薄;真武汤给药后全心变小,左心室腔较模型组减小,室壁相对厚度变大;（3）光镜下左心室心肌HE染色显示：cTnTR141W转基因小鼠心肌排列紊乱,真武汤给药后心肌细胞排列较整齐。（4）透射电镜观察显示,正常组小鼠的心肌纤维排列有序,结构规则,线粒体较多,闰盘平滑而整齐;而模型组小鼠的心肌纤维变细,甚至溶解并消失,线粒体肿胀并呈空泡化,闰盘卷曲而不整齐。中药真武汤干预组心肌纤维的排列、肌小节及线粒体的变化均有改善;（5）HpaⅡ酶切和RT-PCR鉴定显示,模型组转基因cTnTR141W是内源性cTnT的1.13倍,真武汤组转基因cTnTR141W是内源性cTnT的0.58倍。结论：真武汤可降低cTnTR141W转基因扩张型心肌病小鼠cTnTR141W基因表达,同时改善cTnTR141W转基因扩张型心肌病小鼠的心脏结构及心肌细胞的超微结构。
Objective：To study the effect of Zhenwu Decoction on cardiomyocyte structure and cTnTR141W gene expression in family hereditary dilated cardiomyopathy by using cTnTR141W transgenic cardiomyopathy mice. Methods：（ 1）The expression of cTnTR141W in blank and normal mice was detected by RT-PCR and HpaⅡ digestion. The cTnTR141W transgenic mice were randomly divided into model group and Zhenwu Decoction group. After 1 month as the end point of administration,pathological changes of heart structure were observed under light microscope in model group and Zhenwu Decoction group.（3） The ultrastructure of myocardium was observed by transmission electron microscopy（ TEM）.（3）The expression of cTnTR141W transgene in myocardium of mice was identified by HpaⅡ digestion and RT-PCR. Results：（1）The expression of cTnTR141W in normal and transgenic mice was detected by HpaⅡ digestion and RT-PCR. The results showed that the gene of the blank group was not detected after cleavage of cTnTR141W ,and those in the model group were digested. The transgenic cTnTR141W was detected and the transgenic cTnTR141W was 1 time of the endogenous cTnT.（2）Scanning section showed that the heart of the model group was larger than that of the normal group and the left ven-tricular was dilated and the wall became thinner.（3） Light microscopy left ventricular myocardial HE staining showed cTnTR141W transgenic mice myocardium disorder. After administration of Zhenwu Decoction,myocardium was arranged neatly.（4） Transmission electron microscopy showed that the normal group’s myocardial fibers arranged in order,the structure was in order and there were more mitochondria and the intercalated disk was smooth and tidy. The model group’s mice myocardial fiber was thinning,and even dissolved and disappeared. The mitochondria was swelling and then vacant,intercalated coil curl and not neat.（5） HpaⅡ digestion and RT-PCR identification showed that the transgenic cTnTR141W in the model group was 1. 13 times of the endogenous cTnT,and the transgenic cTnTR141W of Zhenwu Decoction group was significantly higher than that of the control group is 0. 58 times of the endogenous cTnT. Conclusion： Zhenwu Decoction can decrease the expression of cTnTR141W gene in cTnTR141W transgenic cardiomyopathy mice and improve the cardiac structure and ultrastructure of cTnTR141W transgenic cardiomyopathy mice.
出处
《中华中医药学刊》
CAS
北大核心
2018年第1期118-121,I0012,共5页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(81173184)
辽宁省科技厅项目(2016010484-301)
